转子相关的TnpB酶的冷EM结构
Ryoya Nakagawa1, Hisato Hirano1, Satoshi N Omura1
1Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan.
Nature
|April 5, 2023
概括
这项研究揭示了TnpB的结构,TnpB是一个微型RNA导向的DNA内核酶,它从转子体演变为CRISPR-Cas12系统. 这为RNA引导的DNA分裂机制提供了洞察力.
科学领域:
- 分子生物学
- 遗传学
- 生物化学
背景情况:
- CRISPR-Cas12效应器属于第二类V型系统.
- 来自IS200/IS605转子超级家族的TnpB蛋白质是微型RNA引导的DNA内核酶.
- TnpB和Cas12的进化链接和DNA裂变机制尚不清楚.
研究的目的:
- 阐明TnpB的结构和机制.
- 了解TnpB和Cas12酶之间的进化关系.
- 研究由TnpB介导的RNA引导的DNA裂变过程.
主要方法:
- 用冷电子显微镜 (cryo-EM) 确定Deinococcus radiodurans ISDra2 TnpB的结构.
- 复杂的形成分析与相关的 ωRNA 和目标DNA.
- 对TnpB活动的功能分析.
主要成果:
- 确定了与 ωRNA 和目标DNA 结合的 TnpB 的冷EM 结构.
- ωRNA形成了一个保存的伪结结构.
- 通过TnpB发现 ωRNA识别和DNA裂变的机制细节.
- 结构比较表明从TnpB到CRISPR-Cas12的进化途径.
结论:
- TnpB使用保存的 ωRNA 伪结来定位DNA.
- 这些发现揭示了CRISPR-Cas12从转子子相关蛋白质的演变.
- 这项研究提供了TnpB功能和CRISPR-Cas演变的机制性见解.
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